Aging can be associated with a decreased ability to respond to metabolic challenges resulting, for example, in fatigue on cognitive tasks or increased susceptibility to substrate deprivation, such as relative hypoglycemia or hypoxia. We hypothesize that cellular and molecular changes in the regulation of intrinsic CNS bioenergetics (i.e., neuronal-glial interactions, aerobic and anaerobic metabolic pathways, substrate availability, etc.) during aging can impair adaptive responses to sustained energy demand. We propose to study prolonged metabolic demand and fatigue in isolated brain tissue from aged animals, in the absence of systemic factors such as poor vasculature or systemic substrate supply, to identify intrinsic changes in neuronal metabolism and neuronal-glial interactions. In vitro brain slices obtained from aged animals retain the in vivo metabolic characteristics of that age, as well as the intrinsic circuits and other factors leading to regulation of metabolism on a local tissue scale. Preliminary experiments indicate that neuronal function and mitochondrial redox state in aging hippocampus are more vulnerable to metabolic stress, such as lowered glucose levels and prolonged synaptic stimulation, compared to tissue from younger animals, suggesting that aged individuals may have reduced ability to support an increased rate of oxidative metabolism for an extended period of time. We will evaluate neuronal fatigue and neuronal-glial interactions during prolonged metabolic stress by studying the energetic relationships between oxygen utilization, mitochondrial redox state, and neuronal activity, using direct tissue lactate, glucose and Po2 measurements, NAD(P)H fluorescence, and neuronal responses in hippocampus. These techniques will be used during prolonged synaptic stimulation (increased metabolic demand) and conditions of limited substrate delivery. These results will facilitate understanding how local tissue responses and bioenergetics affect metabolism in aging. The understanding of the mechanisms underlying neuronal fatigue may indicate novel targets for treatment which may enhance performance on sustained cognitive tasks. PUBLIC HEALTH RELEVANCE: This proposal seeks to understand how metabolism in the brain changes with aging, assessing both mechanisms underlying fatigue to persistent responses and possible treatment directions. The goal is to assess components of oxidative and glycolytic metabolism, particularly during sustained metabolic demand, over minutes, which are intrinsic to neurons and glia. The in vitro slice preparation proposed here allows assessment of local metabolic interactions directly in brain tissue, without direct involvement of the vascular system and systemic provision of substrates.